The Journal of Phytopharmacology 2017; 6(6): 359-363 Online at: www.phytopharmajournal.com

Research Article Pharmacognostic, physicochemical and phytochemical ISSN 2320-480X investigation of maderaspatana JPHYTO 2017; 6(6): 359-363 November- December Tanvi Dodiya*, Dr. Vineet Jain Received: 05-11-2017 Accepted: 11-12-2017 ABSTRACT © 2017, All rights reserved

Grangea maderaspatana is a popular Indian medicinal belonging to the family. This plant is Tanvi Dodiya Parul Institute of Pharmacy & Research, commonly known as Madras Carpet. It has long been used in traditional Ayurvedic medicine for various P.O. Limda, Tal. Waghodia, Dist. diseases. Literature reveals that the phytoconstituents like steroids, terpenes, flavonoids, saponins, Vadodara, Gujarat-391760, India carbohydrates, phenolics and tannins are present in the plant. Pharmacognostical standards on G. maderaspatana are not yet available for correct identification of plant material and to ascertain its quality and Dr. Vineet Jain Bhagwan Mahavir College of Pharmacy, purity. The present study was therefore undertaken to determine the required pharmacognostical standards Surat, Gujarat-395017, India according to the Pharmacopoeial guidelines for evaluation of the plant. The study included examination of morphological, microscopical, physicochemical properties, phytochemical analysis and HPTLC fingerprinting. The standardization parameters presented in this paper may be suggested to establish the authenticity of Grangea maderaspatanaand can also help to distinguish the drug from its other species. The pharmacognostic

profile of the plant presented here will contribute in standardization viz., quality, purity and sample identification.

Keywords: , Asteraceae, Standardization.

INTRODUCTION

Grangea maderaspatana is a popular Indian medicinal plant belonging to the Asteraceae family. This plant is commonly known as Madras Carpet growing in sandy lands and waste places. It is reported to

contain flavonoids, diterepenes, sesquiterpenoids, steroid, and essential oil. The herb is good for pain in

the eyes and ears. The root is an appetizer; astringent to the bowels, diuretic, anthelmintic, emmenogogue, galactogogue, stimulant; useful in griping, in troubles of the chest and lungs, headache, paralysis, rheumatism in the knee joint, piles, pain in the muscles, diseases of the spleen and the liver, the troubles of the ear, the mouth and the nose; lessens perspiration (Unani). The plant is stomachic and uterine stimulant [1].

Figure 1: G. maderaspatana Plant

MATERIALS AND METHODS Correspondence: Tanvi Dodiya Parul Institute of Pharmacy & Research, Plant collection and Identification P.O. Limda, Tal. Waghodia, Dist. Vadodara, Gujarat-391760, India The plant of Grangea maderaspatana was collected in the month of December from Saputara, Gujarat. Email: tanvi.dodiya[at]paruluniversity.ac.in Authentication was done by Taxonomist of the Botanical Survey of India, Jodhpur. A voucher specimen

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(No BSI/AZRC/I.12012/Tech./2015-16/419) was deposited in the RESULTS Herbarium of Botanical Survey of India, Jodhpur. Macroscopic study Macroscopic and microscopic investigation The leaves of Grangea maderaspatana are simple, alternate, sessile, The macroscopic features of Grangea maderaspatana leaf, stem and Oblong-obovate, coarsely dentate, obtuse, reticulate, 3.5-7.5 cm long root was determined using prescribed method2. For microscopical and 1.5-2.5 cm wide. They are dark green in color with characteristic examinations, free hand sections of the fresh leaf, stem and root were odor and taste. The stem is Prostrate, ascending to erect, which is up cut, cleared with chloral hydrate solution and water, and stained with to 55 cm tall, branched from the base. The stem is green colored with a drop of hydrochloric acid and phloroglucinol. Photomicrographic numerous hairs on the surface. Roots are primarily tap-rooted but images were taken by using Trino CXR camera. form weak adventitious roots along the stolon nodes. They are white to brown color with characteristic odor and taste. Leaf constant Flowers: The inflorescence is a terminal, truncate-spherical head, 6- Leaf constants such as stomatal index, vein islet number, vein 10 mm in diameter, solitary or 2-3 together, yellow colored. The termination number and palisade ratio were studied according to peduncle is 1-4 cm long. The flowers are all tubular and about 1.5 mm standard methods [2]. long.

Physicochemical analysis Fruit: The fruit is pale brown and compressed. Achenes are cylindric, glandular, and about 2 millimeters long. Physicochemical analysis, such as extracting values, Loss on drying and ash values were performed according to the official methods [3, 4, Microscopic study 5]. Leaf: The transverse section of leaf of Grangea maderaspatana Preliminary phytochemical screening shows dorsiventral nature. The section is broadly divided in to lamina and midrib region. The lamina of leaf shows three distinct regions Preliminary phytochemical screening was carried out by using viz., upper epidermis, lower epidermis and mesophyll. The upper and standard procedures [6, 7]. lower epidermis is single layer of cells covered by cuticle. The mesophyll is differentiated in to palisade and spongy parenchyma. HPTLC Fingerprinting The palisade parenchyma are narrow, closely packed, elongated cylindrical cells.The cells of spongy parenchyma are compactly HPTLC fingerprinting of chloroform and methanol extracts of G. arranged. Multicellular covering trichomes and glandular trichomes maderaspatanawas performed for oleanolic acid and ursolicacid [8]. are seen. a) Sample preparation: Accurately weighed 20 mg of each Midrib shows the epidermal cells are polygonal in shape covered by extracts individually into volumetric flask and 10 mL methanol thin cuticle. Few collenchymatous cells are present below the upper was added to it. Dissolved it and filtered it with whatman filter epidermis and above the lower epidermis. The ground tissue is paper no. 1 and used for HPTLC profiling. parenchymatous. The vascular bundle is bicollateral type. b) Standard preparation: Accurately weighed 10 mg of each standard individually into volumetric flask and 10 mL methanol was added to it. Dissolved it and filtered it with whatman filter paper no. 1 and used for HPTLC profiling.

Chromatographic Conditions:

Application Mode CAMAG Linomat 5 - Applicator Application of sample Automatic device “CAMAG LINOMAT – 5” Stationary Phase MERCK - TLC / HPTLC Silica gel 60 F254 on Aluminum sheets (10 x 10 cm) Figure 2a: T.S of G. maderaspatana leaf (stained) Application Volume 10 µL Mobile Phase Toluene: Ethyl acetate: Formic acid (8: 2: 0.1) Development Mode CAMAG TLC Twin Trough Chamber Spray reagent Anisaldehydesulphuric acid reagent Derivatization mode CAMAG – Dip tank for about 1 minute Visualization @ 510 nm after derivatization

Figure 2b: T.S of G. maderaspatana leaf showing lamina

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Figure 2c: T.S of G. maderaspatana leaf showing midrib

Stem: Transverse section of stem shows circular outline. The epidermis is single layered covered with cuticle. Epidermis consists of multicellular covering trichomes. Cortex is 6 to 8 layers thick and made up of thin walled parenchymatous cells. Some cells contain microsphenoidal calcium oxalate crystals. Endodermis is indistinct. Figure 4a: T.S. of G. maderaspatana root (unstained) Phloem is narrow zone consisting of 4 to 6 layers of cells. The xylem consists of small vessels and parenchyma. The pith in the centre is large and made up of thin walled parenchymatous cells.

Figure 3a: T.S. of G. maderaspatana stem (unstained)

Figure 4b: T.S. of G.maderaspatana root (stained)

Powder characteristics

The organoleptic evaluation of powder revealed the following characteristics. The powder is light green color with characteristic odor and taste. On microscopic examination, the powder showed lamina, anisocytic and anomocytic stomata, multicellular covering trichomes, lignified fibres, Phloem, Cork and Xylem.

Figure 3b: T.S. of G. maderaspatana stem (stained)

Root: Transverse section of root shows circular outline. Cork is made up of 2-4 layers of thin walled cells. Cortex is made up of 5-7 layers of rectangular parenchymatous cells. The pericycle appers like semilunar patches of pericyclic fibres with parenchyma in between. The Phloem is 5-7 layers thick. The xylem consists of small vessels, Lamina Multicellular covering trichomes fibres,

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of exhausted or adulterated drugs. The results are presented in table no. 2.

Table 2: Physico-Chemical Parameters of powder of Grangea maderaspatana

Sr. No. Parameters Values (% w/w) 1 Loss on drying 8.0% 2 Ash value

Total ash 11.6% Lignified fibres Cork Acid insoluble ash 1.8% Water soluble ash 4.5% 3 Extractive value Water soluble extractive 10.0% Alcohol soluble extractive 10.0%

Preliminary phytochemical screening

Preliminary phytochemical screening revealed presence of carbohydrates, phytosterols, saponins, terpenoids, flavonoids, tannins and phenolics (Table no. 3)

Phloem Stomata Table 3: Phytochemical screening of extracts of Grangea maderaspatana

Chemical Petroleum Chloroform Ethyl Methanol Water constituents ether extract extract acetate extract extract extract Carbohydrate - - - + ++ Protein - - - - - Phenolics & - - - + + Tannins Saponins - - - + ++

Flavanoids - - - + + Xylem Terpenes ++ ++ + + - Steroids ++ ++ + - - (Anisocytic & Anomocytic) Alkaloids - - - - -

Figure 5: Powder characteristics of Grangea maderaspatana powder (+ - Positive test, - - Negative test)

Leaf constants HPTLC Fingerprinting:

The leaf constants viz., stomatal index, vein islet number, vein termination number and palisade ratio are presented in table no. 1.

Table 1: Leaf constants of Grangea maderaspatana leaf

Leaf constants Values Stomatal index 25-33 Vein islet number 2-5 Vein termination number 9-14 1. Gm. MeOH ext. 2. Std – Oleanolic acid Palisade ratio 6.00-8.00 3. Std – Ursolic acid 4. Gm. CHCl ext. 3 Physico-chemical parameters

The ash values of a drug gave an idea of the earthy matter or the inorganic composition and other impurities present along with the Figure 6a: HPTLC plate after derivatization with Anisaldehyde sulphuric acid drug. The extractive values are primarily useful for the determination

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5. Gm. MeOH ext. 6. Std – Oleanolic acid 7. Std – Ursolic acid 8. Gm. CHCl ext. Figure 6b: Chromatogram of standard Oleanolic acid 3 Figure 6f: HPTLC 3DChromatogram

CONCLUSSION

The microscopic character, leaf constants, phytochemical screening and physicochemical constants studied here can be used for judging the adulteration and purity of this drug. As there is no pharmacognostic anatomical work on record for this drug, the present work was taken up with a view to lay down standards which could be useful to detect the adulteration and purity of this plant

REFERENCES

1. Kirtikar K, Basu B. Indian Medicinal . Vol. II, International Book distribution, Kolkata, 2004. 2. Evans WC. Trease and Evans’ Pharmaconosy. 15th Ed. W.B. Sounders Figure 6c: Chromatogram of standard Ursolic acid Company Ltd, London, 2005. 3. Anonymous: World Health Organization. Quality control methods for medicinal plant materials, WHO/PHARM/92.559,1998. 4. Anonymous. The Indian Pharmacopoeia. Vol. II. The Controller of Publications, New Delhi, 1996. 5. Mukherjee PK. Quality Control of Herbal Drugs: An Approach to Evaluation of Botanicals. Business Horizones, 2002. 6. Khandelwal KR. Practical Pharmacognosy Technique and Experiments. 13th Ed, Nirali Prakashan, Pune, 2005. 7. Harborne JB. Phytochemical methods, A guide to modern techniques of plant analysis, 3rd Ed, Chapman and Hall, London, 2007. 8. Gupta M, Bisht D, Khatoon S, Srivastava S. Determination of ursolic acid a biomarker in different Swertia Species through High Performance Thin Layer Chromatography. Chinese Medicine, 2011; 2:121-124.

HOW TO CITE THIS ARTICLE Dodiya T, Jain V. Pharmacognostic, physicochemical and phytochemical investigation of Grangea maderaspatana. J Phytopharmacol 2017; 6(6):359-363. Figure 6d: Chromatogram of chloroform extract of Gm

Figure 6e: Chromatogram of methanol extract of Gm

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